Application of active ingredients of ancient-Chinese health-preserving essence in preparation of medicine for treating and / or preventing vascular dementia

Abstract

The invention provides application of active ingredients of ancient-Chinese health-preserving essence in preparation of a medicine for treating and / or preventing vascular dementia, and belongs to the technical field of biological medicines. The active ingredients of the ancient-Chinese health-preserving essence comprise one or more of 20 active ingredients including deoxyharringtonine, schisantherin and the like. According to the invention, 20 active components and 20 core protein targets of the ancient-Chinese health-preserving essence for treating vascular dementia are obtained through network pharmacological analysis. According to the invention, 20 active components and 20 core protein targets are subjected to molecular docking, and the result shows that the active components and the core protein targets generally have good binding activity. The application prospect of the active ingredients of the ancient-Chinese health-preserving essence in preparation of the medicine for treating and / or preventing the vascular dementia can be defined, and technical support is provided for research, development and screening of the medicine for treating and / or preventing the vascular dementia.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to the use of active ingredients of Gu Han Health Essence in preparing medicines for treating and / or preventing vascular dementia. Background Art

[0002] Gu Han Yang Sheng Jing is a traditional Chinese medicine compound preparation derived from traditional Chinese medicine theory. It is primarily used to tonify Qi, nourish the kidneys, and improve essence. It is suitable for symptoms such as Qi and blood deficiency, dizziness, palpitations, fatigue, and weakness. Its core ingredients include ginseng, astragalus, wolfberry, polygonatum, and Ligustrum lucidum. However, the mechanism of action of Gu Han Yang Sheng Jing in treating vascular dementia is currently unclear, and the active ingredients in Gu Han Yang Sheng Jing for this treatment remain uncertain. Summary of the Invention

[0003] The purpose of the present invention is to provide the use of the active ingredients of Gu Han Health Essence in the preparation of a drug for treating and / or preventing vascular dementia, thereby solving the current technical problem that the active ingredients of Gu Han Health Essence for treating vascular dementia are unclear.

[0004] The present invention provides an application of the active ingredients of Gu Han Health Essence in the preparation of a medicine for treating and / or preventing vascular dementia. The active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochrom eno[3,4-h]isoquinolin-2-ium, 1-methoxyphanetin, 4'-methoxyglabridin, α-tocopherylquinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-trimethoxyclitanane, 3'-hydroxy-4'-O-methylglabridin, ginsenoside Rh4, glabridin, benzanoside, tricholizine B, neoclidinone, baicalein, succinolide, meditartin, and vesin.

[0005] Preferably, the active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester E, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin, 4'-methoxyglabridin, α-tocopheryl quinone and licorice chalcone.

[0006] Preferably, the active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin and 4'-methoxyglabridin.

[0007] Preferably, the active ingredients of the Gu Han Health Essence include deoxyharringtonine and / or schisandra chinensis monohydrate.

[0008] Preferably, the active ingredients of the Gu Han Health Essence treat and / or prevent vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1, NOS2, PIK3CA, PIK3CD, GSK3B, RAF1, CDK2, PIK3CB, KDR, ADORA2A, SRC, ADORA1, PTGS2 and BACE1.

[0009] Preferably, the active ingredients of the Gu Han Health Essence achieve the treatment and / or prevention of vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1 and BACE1.

[0010] Preferably, the medicine uses Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, the medicine uses the active ingredient of Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, in addition to the active ingredient of Gu Han Health Essence, the medicine also includes other active ingredients for treating and / or preventing vascular dementia.

[0011] Preferably, the dosage form of the drug includes an oral dosage form.

[0012] Preferably, the oral dosage form includes solid preparations or liquid preparations; solid preparations include powders, tablets, granules, capsules or pills; and the liquid preparations include mixtures, oral liquids or decoctions.

[0013] The present invention also provides the use of the active ingredients of Gu Han Health Essence in the screening and / or development of vascular dementia drugs, wherein the active ingredients of Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromene no[3,4-h]isoquinolin-2-ium, 1-methoxyphanethorn, 4'-methoxyglabridin, α-tocopherylquinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-trimethoxyclitanane, 3'-hydroxy-4'-O-methylglabridin, ginsenoside Rh4, glabridin, benzanoside, tricholizine B, neoclidinone, baicalein, succinolide, meditartin, and vesin.

[0014] The present invention provides an application of the active ingredients of Gu Han Health Essence in preparing a medicine for treating and / or preventing vascular dementia. The active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxy-phaseolin, 4'- Methoxyglabridin, α-tocopherylquinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-trimethoxypterocarpan, 3'-hydroxy-4'-O-methylglabridin, ginsenoside Rh4, glabridin, benzaloline, tricholizine B, shinpterocarpin, baicalein, succinolide, meditartin, and vesin. The present invention identified 20 active ingredients and 20 core protein targets for Gu Han Yang Sheng Jing in the treatment of vascular dementia through network pharmacology analysis. Molecular docking of the 20 active ingredients with the 20 core protein targets revealed generally good binding activity between the 20 active ingredients and the 20 core protein targets. The average docking score of the dementia-related protein target BACE1 with 20 compounds was -7.845 kcal / mol, which also showed good binding activity. This can clarify the application prospects of the active ingredients of Gu Han Health Essence in the preparation of drugs for the treatment and / or prevention of vascular dementia, and also provide technical support for the development and screening of drugs for the treatment and / or prevention of vascular dementia. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0016] Figure 1 is the intersection target Venn diagram;

[0017] Figure 2 For PPI network;

[0018] Figure 3 It is the GO molecular function map;

[0019] Figure 4 It is the GO biological process diagram;

[0020] Figure 5 This is the GO cell composition diagram;

[0021] Figure 6 It is the KEGG pathway map;

[0022] Figure 7 It is a compound-target-pathway network diagram;

[0023] Figure 8 is the molecular docking heat map;

[0024] Figures 9 - 11 Schematic diagram of 2D-3D molecular docking; among them, a1: RS11-ESR1; a2: RS11-NOS2; a3: RS11-PIK3CA; b1: RS14-ESR1; b2: RS14-EGFR; b3: RS14-NOS2; b4: RS14-PIK3CA; b5: RS14-BACE1; c1: YYH2-NOS2; c2: YYH2-PIK3CA; c3: YYH2-BACE1; d1: GC43-EGFR; d2: GC43-NOS2; d3: GC43-PIK3CA; d4: GC43-BACE1; e1: GC47-ESR1; e2: GC47-EGFR; e3: GC47-NOS2. DETAILED DESCRIPTION

[0025] The present invention provides an application of an active ingredient of Gu Han Health Essence in preparing a medicine for treating and / or preventing vascular dementia. The active ingredient of Gu Han Health Essence comprises one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinol in-2-ium, 1-methoxyphanethorin, 4'-methoxyglabridin, α-tocopherylquinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-Trimethoxypterocarpan, 3'-Hydroxy-4'-O-Methylglabridin, ginsenoside Rh4, glabridin, benzylospermine, tricholizine B, Shinpterocarpin, baicalein, succinolide, meditarostane, and vesicin.

[0026] In the present invention, the 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, molecular number MOL004388, has a molecular weight of 60.64, a molar mass of 60.64 g / mol, and a molecular formula of C 17 H 18 NO4.

[0027] In the present invention, the chemical structure of the 3,9,10-Trimethoxypterocarpan is (6aR,11aR)-3,9,10-trimethoxy-6a,11-dihydro-6H-[1,3]benzofurano[3,2-c]chromene, and the molecular formula is C 18 H 18 O5, molecular weight is 306.35.

[0028] In the present invention, the Shinpterocarpin belongs to the isoflavone compound, and its molecular formula is C 20 H 18 O4, molecular weight is 322.36 g / mol, CAS registration number is 157414-04-5.

[0029] In the present invention, the molecular formula of the 3'-Hydroxy-4'-O-Methylglabridin (3'-hydroxy-4'-O-methylglabridin is C 21 H 22O5, molecular weight is 354.3964.

[0030] As an embodiment, the active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester E, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin, 4'-methoxyglabridin, α-tocopheryl quinone and licorice chalcone.

[0031] As an embodiment, the active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin and 4'-methoxyglabridin.

[0032] As an embodiment, the active ingredients of the Gu Han Health Essence include deoxyharringtonine and / or schisandra chinensis ester B.

[0033] As an embodiment, the active ingredients of the Gu Han Health Essence treat and / or prevent vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1, NOS2, PIK3CA, PIK3CD, GSK3B, RAF1, CDK2, PIK3CB, KDR, ADORA2A, SRC, ADORA1, PTGS2 and BACE1.

[0034] As an embodiment, the active ingredients of the Gu Han Health Essence treat and / or prevent vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1 and BACE1.

[0035] The present invention conducts molecular docking between the above 20 active ingredients and 20 protein targets. The results show that the 20 active ingredients and the 20 protein targets generally have good binding activity, for example, deoxyharringtonine (YYH2)-NOS2, schisandra chinensis ester ethyl (RS11)-PIK3CA, 1-methoxyphaseolin (HQ3)-PTGS2, etc.

[0036] As an embodiment, the drug uses Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, the drug uses the active ingredient of Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, in addition to the active ingredient of Gu Han Health Essence, the drug also includes other active ingredients for treating and / or preventing vascular dementia.

[0037] As an embodiment, the dosage form of the drug includes an oral dosage form.

[0038] As an embodiment, the oral dosage form includes a solid preparation or a liquid preparation; the solid preparation includes a powder, tablet, granule, capsule or pill; the liquid preparation includes a mixture, oral liquid or decoction.

[0039] The present invention also provides the use of the active ingredients of Gu Han Health Essence in the screening and / or development of vascular dementia drugs, wherein the active ingredients of Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ester ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinoli n-2-ium, 1-methoxyphanethorin, 4'-methoxyglabridin, α-tocopherylquinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-Trimethoxypterocarpan, 3'-Hydroxy-4'-O-Methylglabridin, ginsenoside Rh4, glabridin, benzylospermine, tricholizine B, Shinpterocarpin, baicalein, succinolide, meditarostane, and vesicin.

[0040] As an embodiment, the active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin, 4'-methoxyglabridin, α-tocopheryl quinone and licorice chalcone; further, the Gu Han Health Essence The active ingredients of Han Yangsheng Jing include one or more of the following ingredients: deoxyharringtonine, schisandrin ester B, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin and 4'-methoxyglabridin; further, the active ingredients of the Gu Han Yangsheng Jing include deoxyharringtonine and / or schisandrin ester B.

[0041] To further illustrate the present invention, the application of the active ingredients of the Gu Han Health Essence provided by the present invention in the preparation of drugs for treating and / or preventing vascular dementia is described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0042] Example 1 Study on the mechanism of action of Gu Han Yang Sheng Jing on vascular dementia (VD) based on network pharmacology

[0043] 1.1 Methods and Materials

[0044] 1.1.1 VD-related target screening

[0045] To collect VD-related targets, the keyword "vascular dementia" was searched in the GeneCards database (version 5.15). The retrieved VD target information (full name and abbreviation) was saved for future use.

[0046] 1.1.2 Construction of the Ancient Chinese Health Essence Ingredient Database

[0047] The TCMSP database was used to search for the active ingredients of Gu Han Yang Sheng Jing.

[0048] Constructing a target database requires predicting and screening the ADME properties of TCM ingredients. ADME refers to the four necessary pathways for drugs to exert their effects: absorption, distribution, metabolism, and excretion. Oral bioavailability (OB), drug-likeness (DL), and blood-brain barrier (BBB) ​​are key ADME parameters. Based on the screening results of ADME parameters, the drug ingredients that are ultimately used for analysis and are able to enter the pathological area and exert their effects are screened. Based on the limiting conditions of OB ≥ 30%, DL ≥ 0.18, and BBB > -0.30, a preliminary screening of active ingredients is performed to obtain active compounds and their targets. After screening, the standardization of protein targets is verified in the Uniprot protein database. After constructing a TCM ingredient database and performing ADME screening on the ingredients, all the brain-entering ingredients of the Gu Han Yangsheng Jing compound were obtained.

[0049] 1.1.3 Construction and analysis of the interaction network between Gu Han Yang Sheng Jing and VD protein

[0050] To investigate the interactions between Gu Han Yang Sheng Jing drug targets and VD targets, we extracted intersecting targets using the Jvenn platform and constructed a Venn diagram. We then uploaded these intersecting targets to the STRING 11.0 database to construct a protein-protein interaction network (PPI) model. After submitting the data, we set the biological species to "Homo sapiens" and the minimum interaction threshold to "high confidence (0.700)" to generate a PPI network. The PPI network was further analyzed using the MCODE plugin in CytoScape 3.7.1 to identify potential protein functional modules.

[0051] 1.1.4 Gene function (GO) and pathway (KEGG) enrichment analysis

[0052] The targets of Gu Han Yang Sheng Jing for treating VD were entered into the Metascape platform, with P < 0.01 set, and functional annotations including cellular components, molecular functions and biological processes and KEGG enrichment analysis were performed. The enriched pathways were analyzed with the help of the Kyoto Encyclopedia of Genes and Genomes (KEGG database), and the data results were visualized using the "Microbiology Information" platform.

[0053] 1.2 Research Results

[0054] 1.2.1 Acquisition of vascular dementia-related targets

[0055] A total of 3,828 VD targets were found in the Genecards database. To focus on highly likely disease targets, targets with a score greater than the median were designated as potential VD targets. The maximum score was 47.18, the minimum was 0.78, and the median was 1.27. Targets with a score > 1.27 were designated as potential VD targets, ultimately yielding 1,500 VD-associated targets.

[0056] 1.2.2 Construction of the Ancient Chinese Health Essence Target Database

[0057] The effective chemical components and targets (without deduplication) of eleven ingredients of Gu Han Yang Sheng Jing, namely, ginseng, astragalus, wolfberry, glossy privet, dodder seed, golden cherry fruit, epimedium, white peony root, malt, polygonatum, and liquorice, were screened with the limiting conditions of OB ≥ 30%, DL ≥ 0.18, and BBB > -0.30. The overview is listed in Table 1.

[0058] Table 1. Effective chemical components and target components of Gu Han Yang Sheng Jing medicinal ingredients

[0059]

[0060]

[0061] The details of the effective chemical components are listed in Tables 2 to 12:

[0062] Table 2 Effective chemical components of ginseng

[0063] MolID MoleculeName OB(%) BBB DL MOL005314 Celabenzine 101.88 0.05 0.49 MOL005308 Aposiopolamine 66.65 0.4 0.22 MOL005321 65.9 0.46 0.34 Frutinone A MOL003648 65.83 0.36 0.54 Inermin MOL005356 61.22 1.22 0.31 Girinimbin MOL000787 59.26 -0.13 0.83 Fumarine MOL005360 57.71 -0.17 0.63 malkangunin MOL005384 57.52 0.28 0.56 suchilactone MOL005320 45.57 0.58 0.2 arachidonate MOL000449 43.83 1 0.76 Stigmasterol MOL002879 43.59 0.26 0.39 Diop MOL005401 39.56 0.21 0.79 ginsenoside Rg5_qt MOL005317 39.27 -0.25 0.81 Deoxyharringtonine MOL000358 36.91 0.99 0.75 beta - sitosterol MOL005399 36.91 0.88 0.75 alexandrin_qt MOL005376 33.09 0.23 0.79 Panaxadiol MOL005357 31.99 0.18 0.83 Gomisin B MOL005348 31.11 -0.18 0.78

[0064] Table 2 Screening of effective targets for all components totals 1094.

[0065] Table 3 Effective chemical components of Astragalus

[0066]

[0067]

[0068] Table 3 Screening of effective targets for all components totals 838.

[0069] Table 4 Effective chemical components of licorice

[0070]

[0071]

[0072] Table 4 Screening of effective targets for all components totals 4285.

[0073] Table 5 Effective chemical components of Epimedium

[0074]

[0075] Table 5 Screening of effective targets for all components totals 663.

[0076] Table 6 Effective chemical components of Polygonatum sibiricum

[0077]

[0078] Table 6 Screening of effective targets for all components totals 553.

[0079] Table 7 Effective chemical components of Ligustrum lucidum

[0080] Ginsenoside - Rh4_qt MolID MoleculeName OB(%) BBB DL MOL000358 36.91 0.99 0.75 beta - sitosterol MOL005169 40.23 0.58 0.82

[0081] Table 7 Screening of effective targets for all components totals 70.

[0082] Table 8 Effective chemical components of Cuscuta australis

[0083]

[0084]

[0085] Table 8 screened a total of 344 effective targets for all components.

[0086] Table 9 Effective chemical components of Rosa laevigata

[0087] (20S)-24 - ene - 3β,20 - diol - 3 - acetate MolID MoleculeName OB(%) BBB DL MOL008628 53.43 0.87 0.26 4'-Methyl - N - methylcoclaurine MOL008622 42.88 0.2 0.82 Methyltrametenolate MOL001494 42 1.14 0.19 Mandenol MOL000358 36.91 0.99 0.75 beta - sitosterol MOL005030 30.7 0.8 0.2

[0088] Table 9 screened a total of 332 effective targets for all components.

[0089] Table 10 Active chemical components of white peony root

[0090] gondoic acid MolID MoleculeName OB(%) BBB DL MOL000211 55.38 0.22 0.78 Mairin MOL000358 36.91 0.99 0.75 beta - sitosterol MOL000359 36.91 0.87 0.75

[0091] Table 10 Screening of effective targets for all components totals 138.

[0092] Table 11 Effective chemical components of wolfberry

[0093]

[0094]

[0095] Table 11 Screening of effective targets for all components totals 1,300.

[0096] Table 12 Effective chemical components of malt

[0097] Sitosterol MolID MoleculeName OB(%) BBB DL MOL010857 57.16 -0.09 0.68 Sterigmato - cystin MOL010861 45.66 1.01 0.48 Delta - D MOL000358 36.91 0.99 0.75 beta - sitosterol MOL000359 36.91 0.87 0.75 sitosterol MOL010862 35.91 -0.01 0.5

[0098] Table 12 Screening of effective targets for all components totals 307.

[0099] 1.2.3 Screening of targets for the intervention of VD with Gu Han Health Essence

[0100] The targets of the screened Gu Han Yang Sheng Jing active ingredients were intersected with the VD disease targets, and a Venn diagram was drawn using the Jvenn platform, resulting in 330 common targets of Gu Han Yang Sheng Jing ingredients and vascular dementia ( α - tocopherylquinone ).

[0101] 1.2.4 Construction and analysis of protein interaction networks

[0102] Submit the target to the STRING11.0 platform to obtain the intersection target PPI network. Figure 1 Since the interactions between proteins in a PPI network are mutual, it is usually classified as an undirected graph. The darker the color and the larger the shape, the higher the density of the protein in the network.

[0103] 1.2.5 Gene function and pathway enrichment analysis

[0104] The Metascape data platform was used to analyze signaling pathways related to Gu Han Yang Sheng Jing's targets, and the results were visualized using Wei Sheng Xin. The results showed that the compound treats VD through multiple targets and pathways. Figure 2 As shown in Tables 13 to 15, the molecular functions related to the treatment of vascular dementia with Gu Han Yang Sheng Jing include protein kinase activity, protein tyrosine kinase activity, kinase binding, neurotransmitter receptor activity, nuclear receptor activity, etc.; the biological processes mainly include the cell's response to nitrogen-containing compounds, response to hormones, positive regulation of phosphorylation, positive regulation of cell migration, response to inorganic substances, etc.; the cellular components mainly include membrane rafts, dendrites, postsynapses, receptor complexes, cytoplasmic perinuclear areas, distal axons, etc. Figures Table 16 shows the KEGG pathway analysis results, which show the top 20 signaling pathways for the treatment of vascular dementia with Gu Han Yang Sheng Jing. The key pathways include cancer pathway, lipid and arteriosclerosis pathway, PI3K-AKT signaling pathway, cAMP signaling pathway, MAPK signaling pathway, Alzheimer pathway, etc.

[0105] Table 13 Molecular function table

[0106]

[0107] Table 14 Biological process table

[0108]

[0109] Table 15 Cell composition

[0110]

[0111]

[0112] Table 16 KEGG pathway table

[0113]

[0114]

[0115]

[0116]

[0117] 1.2.6 Construction and analysis of the target network of Gu Han health essence intervention for VD

[0118] The ancient Chinese health essence component-vascular dementia target-pathway network was constructed using CytoScape3.7.1. ​ and Tables 17 and 18 (darker colors and larger shapes in the figures represent more important roles within the network). Using the built-in Network Analyzer in CytoScape 3.7.1, the network topology parameters of the Gu Han Yang Sheng Jing treatment for VD were analyzed to identify core components and targets. The results show that the active components with the highest degree values ​​in this network are RS11, RS14, YYH2, GC43, and GC47, and the target proteins with the highest degree values ​​include ESR1, ESR2, AR, EGFR, HSP90AA1, NOS2, PIK3CA, PIK3CD, GSK3B, and RAF1, among others.

[0119] Table 17 Network node characteristic parameters of the main active ingredients of Gu Han Health Essence

[0120] ​ ​ ​ ​ ​ ​ 82 2.123 0.055 0.471 ​ 80 2.144 0.050 0.466 ​ 71 2.155 0.055 0.464 ​ 54 2.226 0.022 0.449 ​ 51 2.275 0.032 0.440 ​ 48 2.308 0.023 0.433 ​ 47 2.210 0.016 0.453 ​ 44 2.243 0.012 0.446 ​ 43 2.308 0.013 0.433 ​ 43 2.281 0.012 0.438 ​ 42 2.346 0.011 0.426 ​ 42 2.243 0.012 0.446 ​ 42 2.302 0.010 0.434 ​ 41 2.351 0.013 0.425 ​ 39 2.319 0.011 0.431 ​ 39 2.444 0.013 0.409 PTGS2 38 2.357 0.021 0.424 BACE1 38 2.477 0.011 0.404 BRAF 36 2.384 0.008 0.419 MTOR 36 2.341 0.007 0.427 MAP2K1 36 2.302 0.008 0.434 CHRM2 36 2.542 0.012 0.393 MAPK14 34 2.335 0.010 0.428 AKT1 33 2.292 0.009 0.436 PPARG 33 2.422 0.011 0.413 RET 33 2.406 0.006 0.416 ABL1 33 2.477 0.005 0.404 JAK2 32 2.341 0.008 0.427 MAPK1 32 2.302 0.007 0.434 MAPK3 32 2.324 0.011 0.430

[0121] Table 18. Feature parameters of the main target network nodes of Gu Han Health Essence

[0122]

[0123]

[0124] 1.2.7 Summary and Discussion

[0125] Network pharmacology analysis revealed that the main active ingredients in Gu Han Yang Sheng Jing for treating VD are deoxyharringtonine, schisandrin ethyl, 1-methoxyphanethiazine, and 4'-methoxyglabridin. Ingredients with the highest node counts are primarily derived from ingredients such as ginseng, licorice, and malt. Enrichment analysis revealed that the targets of interest are primarily involved in pathways such as cancer pathways, lipid and atherosclerosis pathways, PI3K-AKT signaling pathways, cAMP signaling pathways, MAPK signaling pathways, and Alzheimer's pathways.

[0126] 1.3 Molecular docking

[0127] After completing the network pharmacology work, we have identified important small molecules (numbered RS11, RS14, YYH2, GC43, GC47, etc.), important pathways, and their key proteins. In the next molecular docking step, we will verify the binding ability of small molecules with proteins. We will select the top 20 proteins and small molecules by node number, perform batch molecular docking to calculate binding energies, and select key combinations to use visualization methods to study the types of interaction forces, hoping to find active ingredients and targets with better binding activity.

[0128] 1.3.1 Preparation of compound molecules

[0129] The first step in molecular docking is to obtain the three-dimensional structure of the desired molecule. The top twenty proteins and small molecules ranked by degree in the compound-target-pathway network were selected and their three-dimensional structures were obtained. For ligand small molecules, PubChem was used to obtain the molecule's 3D structure sdf file. For molecules without a 3D structure in the database (usually due to excessively long saturated side chains and numerous conformations, resulting in no determined structure), their 2D structure sdf files were downloaded. Openbabel, a molecular structure file format conversion tool, was used to convert the sdf file to a pdb format file, and the 2D molecular structure was also converted to a 3D structure. Autodocktools was then used to set the single bonds for free rotation of all small molecules one by one. For protein macromolecules, the PDB database was searched for the proteins required in the network pharmacology results. The cif files of the protein structures with the highest scores were selected, and the database code for the selected structures was recorded. Pymol was used to perform desolvation, counterion removal, and hydrogenation on the protein structure. After hydrogenation, Autodocktools was used to set the docking box parameters. When setting up the docking box, it is required that the docking box can basically cover the entire protein macromolecule. This will increase the computational workload, but it will ensure the reliability of the results. After completing the hydrogenation and setting up the docking box, the structure is saved in pdbqt format, and the docking box parameters are saved in txt format for future use.

[0130] 1.3.2 Run the docking program

[0131] Molecular docking calculations were performed using the program Quickvina-W. After preparing the protein and small molecule structures for docking, we batch-called Quickvina-W for docking. The binding energies (affinity, in kcal / mol) generated by Quickvina were recorded and saved as pdbgt files for the docked conformations.

[0132] 1.3.3 Run the docking program

[0133] After confirming the docked conformation, further calculations can be performed to reveal the specific details of the interaction between the small molecule and the protein, such as the protein groups surrounding the small molecule, the spatial conformation of the small molecule, and the specific types of forces acting between the protein and the small molecule. Discovery Studio 2021 will be used to render 2D force maps and 3D spatial environment images. Due to the large number of results obtained from batch docking, it is difficult to render force maps for each. Based on the docking results and the conclusions from the network pharmacology section, force diagrams will be drawn for selected combinations.

[0134] 1.4 Research Results

[0135] 1.4.1 Molecular docking heat map

[0136] The heat map drawing results are as follows Figure 8 The labels range from -4 to -11, with units in kcal / mol. The two ends are colored blue and red, respectively, and the middle region around -7 is colored white. Generally speaking, a molecular docking binding energy less than -5 kcal / mol is considered to have moderate docking activity, while a binding energy less than -7.5 kcal / mol is considered to have relatively good docking activity. As can be seen, the protein-small molecule combinations corresponding to the red grid points in the figure have relatively good docking activity. Molecular docking of 20 core potential compounds with 20 core targets yielded 400 receptor-ligand docking results. Based on the above criteria, 381 showed moderate activity and 308 showed good activity. This suggests that the potential core compounds screened may generally have good binding activity with the core targets. The average docking score for the 20 compounds, BACE1, a protein target associated with dementia, was -7.845 kcal / mol, also indicating good binding activity. These docking results provide technical support for subsequent drug screening.

[0137] 1.4.2 Molecular docking 2D diagram

[0138] Based on the conclusions of network pharmacology and molecular docking results, we selected the top five molecules (RS11, RS14, YYH2, GC43, GC47) and five proteins with good docking results and involved in important pathways, namely ESR1, EGFR, NOS2, PIK3CA, and BACE1. We selected some favorable conformations for each of them and used Discovery Studio 2021 to draw 2D force and 3D spatial environment images ( Figures 9 to 11 ).

[0139] The upper half of each figure above is a 2D force diagram, with different color labels representing different force types (see the legend in the lower left corner of each figure). The bright green labels are conventional hydrogen bonds, which are the forces of greater concern. The lower half of each figure is a 3D spatial environment image, in which the membrane diagram shows the tendency of molecules to form hydrogen bonds. Pink indicates that the area is prone to donating polar hydrogen to form hydrogen bonds (donor), and green indicates that the area is prone to accepting polar hydrogen to form hydrogen bonds (acceptor).

[0140] 1.4.3 Summary and Discussion

[0141] As can be seen from the heat map, with the exception of SRC and MTOR, the other proteins exhibited relatively good binding affinity for most small molecules in molecular docking. The median binding energy for the 400 docking combinations was around -7, indicating that approximately half of the combinations had good binding activity, preliminarily validating the significant therapeutic effect of Gu Han Yang Sheng Jing on vitamin D deficiency. Combinations such as RS11-ESR1, RS11-NOS2, RS11-PIK3CA, RS14-BACE1, YYH2-PIK3CA, and GC43-BACE1 all exhibited at least one conventional hydrogen bond, indicating plausible binding conformations. These results suggest that these combinations are likely gateways for drug compounds to enter the protein pathway and exert their effects.

[0142] In summary, the present invention uses network pharmacology and molecular docking methods to study the mechanism of action of Gu Han Yang Sheng Jing on vascular dementia. By screening the intersection targets of Gu Han Yang Sheng Jing ingredients and vascular dementia, as well as various means such as enrichment analysis, network construction, and molecular docking, a preliminary analysis of the mechanism of Gu Han Yang Sheng Jing's action on vascular dementia was made from the perspective of systems biology. The analysis results showed that ingredients such as deoxyharringtonine, schisandrin, glycyrrhizin and its derivatives, and 1-methoxyphaseolin had the highest correlation in the network and had the highest activity. The pathways of drug action mainly include the PI3K-AKT signaling pathway, the cAMP signaling pathway, the MAPK signaling pathway, and the Alzheimer's pathway, and the main proteins involved are ESR1, EGFR, NOS2, PIK3CA, BACE1, etc. The activation of neural stem cells and the inhibition of neuronal apoptosis are the main mechanisms of action of these pathways.

[0143] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. Use of the active ingredient of Gu Han Health Essence in the preparation of a medicament for treating and / or preventing vascular dementia, characterized in that: The active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester B, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphanethizoside, 4'-methoxyglabridin, α-tocopheryl quinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-trimethoxyclitanane, 3'-hydroxy-4'-O-methylglabridin, ginsenoside Rh4, glabridin, benzylsuccinate, tricholoma B, neoclitan, baicalein, succinolide, meditartin and viscerin.

2. The use according to claim 1, characterized in that The active ingredients of the Gu Han health essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester E, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin, 4'-methoxyglabridin, α-tocopheryl quinone and licorice chalcone.

3. The use according to claim 1, characterized in that The active ingredients of the Gu Han health essence include one or more of the following ingredients: deoxyharringtonine, schisandrin ethyl, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphaseolin and 4'-methoxyglabridin.

4. The use according to claim 1, characterized in that The active ingredients of the Gu Han Health Essence include deoxyharringtonine and / or schisandra chinensis ester B.

5. The use according to any one of claims 1 to 4, characterized in that: The active ingredients of the Gu Han Health Essence treat and / or prevent vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1, NOS2, PIK3CA, PIK3CD, GSK3B, RAF1, CDK2, PIK3CB, KDR, ADORA2A, SRC, ADORA1, PTGS2 and BACE1.

6. The use according to claim 5, characterized in that The active ingredients of the Gu Han Health Essence treat and / or prevent vascular dementia by acting on one or more of the following protein targets: ESR1, ESR2, AR, EGFR, HSP90AA1 and BACE1.

7. The use according to any one of claims 1 to 4, characterized in that The medicine uses Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, the medicine uses the active ingredient of Gu Han Health Essence as the sole active ingredient for treating and / or preventing vascular dementia; or, in addition to the active ingredient of Gu Han Health Essence, the medicine also includes other active ingredients for treating and / or preventing vascular dementia.

8. The use according to any one of claims 1 to 4, characterized in that: The dosage form of the drug includes an oral dosage form.

9. The use according to claim 8, characterized in that The oral dosage forms include solid preparations or liquid preparations; solid preparations include powders, tablets, granules, capsules or pills; and liquid preparations include mixtures, oral liquids or decoctions.

10. The application of the active ingredients of Gu Han Health Essence in the screening and / or development of drugs for vascular dementia is characterized by: The active ingredients of the Gu Han Health Essence include one or more of the following ingredients: deoxyharringtonine, schisandra chinensis ester B, 6-Hydroxy-11,12-dimethoxy-2,2-dimethyl-1,8-dioxo-2,3,4,8-tetrahydro-1H-isochromeno[3,4-h]isoquinolin-2-ium, 1-methoxyphanethizoside, 4'-methoxyglabridin, α-tocopheryl quinone, licorice chalcone, 3'-methoxyglabridin, 3,9,10-trimethoxyclitanane, 3'-hydroxy-4'-O-methylglabridin, ginsenoside Rh4, glabridin, benzylsuccinate, tricholoma B, neoclitan, baicalein, succinolide, meditartin and viscerin.

Images